A multicolor biosensor for alkaline phosphatase activity detection based on the peroxidase activity of copper nanoclusters and etching of gold nanorods

Abstract

Alkaline phosphatase (ALP) plays a vital role in clinical diagnoses and biomedical research. It is important to develop some convenient but sensitive methods for ALP activity detection. In this study, a multicolor biosensor for ALP activity has been developed based on the peroxidase activity of copper nanoclusters (CuNCs) and etching of gold nanorods (AuNRs). The presence of CuNCs can catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) to produce blue TMB⁺. In acid solution, TMB⁺ can cause the etching of AuNRs accompanied by a significant color change of the system. The presence of sodium pyrophosphate (PPi) can inhibit the peroxidase activity of CuNCs, which can be recovered after the addition of ALP. Different ALP added results in the recovery of the catalytic activity of CuNCs to different degrees and generates different amounts of TMB⁺. This consequently affected the morphology of the AuNRs in the system and results in the output of a vivid color change, which can be recognized with the naked eyes easily without any complicated instruments. The biosensor has a linear relationship with ALP activity in the range of 10.0–80.0 U L⁻¹, and the detection limit is 4.6 U L⁻¹. The proposed method has been applied to detect ALP activity in human serum samples with satisfactory results.